Analysis Of Mammalian Metaphase Chromosomal RNA Component

In the universe of science, two worlds have recently collided-those of RNA and chromatin. The intersection of these two fields has been impending, but evidence for such a meaningful collision has only recently become apparent.Metaphase chromosome is perhaps the most accessible large scale structural biology project, in comparison to other chromosomal states. The significance is that this is the most visible biological structure (the famous X), yet we virtually have no idea how it is folded and how these can be changes. RNA interference-mediated heterochromation assembly and gene silencing, So far, there is no report on the identity of the RNAs involved. It is an interesting question whether small RNAs are present or required to form the compact structure. It is now shown that heterochromatin may require some transcription that is coupled to small RNA. So this research will provide very important direction and reference value for metaphase chromosome structure and epigenetics. In my paper, I purified metaphase chromosomes from human HT1080 cell line then isolate RNA components from them, see if there's small RNA fragments that have relationship with metaphase chromosome and transcription regulation exist, and set up a small amount microRNA cloning method, tried to analysis its sequences.Finally I discovered chromosomal RNAs mainly contain ribosomal 28S,18S and 5S subunits, and, I found there is a small band between 20-30bp exsits when detected on 15% polyacrylamide TBE-urea gel.This paper has set up a series of technologies, like metaphase chromosomal isolation method, chromosome in fluorescence situ hybridization, chromosomal RNA isolation method and 3 pmol small initial amount microRNA cloning method by a special molecular cloning procedure based on Weissman' science paper in 2009, so current results provide a good condition for the future analysis and identification of small RNAs sequences from metaphase chromosome.